Western blotting analysis using anti-EIF2AK2 antibody (Cat#63491). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-EIF2AK2 antibody (Cat#63491, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226). EIF2AK2, eukaryotic translation initiation factor 2 alpha kinase 2.
Western blotting analysis using anti-EIF2AK2 antibody (Cat#63491). EIF2AK2 expression in wild type (WT) and EIF2AK2 shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with anti-EIF2AK2 antibody (Cat#63491, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Validation of EIF2AK2 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with anti-EIF2AK2 antibody (Cat#63491, 1:2,000) and analyzed using BD flow cytometer.
Immunocytochemical staining of HeLa cells using anti-EIF2AK2 antibody (Cat#63491, 1:1,000), Top panel: wild-type (WT); Bottom panal: EIF2AK2 shRNA knockdown (KD). Nuclei were stained blue with DAPI; EIF2AK2 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.